伊洛前列素对ILC2s介导的小鼠急性过敏性气道炎症的作用研究

Effects of iloprost on ILC2s in a mouse model of acute allergic airway inflammation

目的:探讨伊洛前列素(Iloprost)对IL-33诱导的小鼠急性过敏性气道炎症中2型固有淋巴细胞(ILC2s)的作用。方法:C57BL/6小鼠随机分为DMSO对照组、IL-33刺激组、IL-33+iloprost干预组和单独iloprost组。HE染色观察肺组织中炎性细胞浸润情况,PAS染色观察黏液分泌情况,流式细胞术检测小鼠支气管肺泡灌洗液(BALF)以及肺组织中嗜酸性粒细胞(EOS)和ILC2s的数量,real-time PCR检测IL-5、IL-13、GATA3和ST2 mRNA的表达量,酶联免疫吸附试验(ELISA)检测BALF和肺匀浆上清中IL-5和IL-13的含量。结果:(1)IL-33刺激组小鼠肺组织切片可见大量炎性细胞浸润且黏液分泌增加;同时,小鼠BALF和肺组织EOS和ILC2s的数量、细胞因子分泌量和mRNA相对表达量均显著高于DMSO对照组和单独iloprost组,差异有统计学意义(P<0.05)。(2)IL-33+iloprost干预组小鼠肺组织切片中炎性细胞浸润程度明显减轻且黏液分泌减少;同时,小鼠BALF和肺组织中EOS和ILC2s的数量、细胞因子分泌量和mRNA相对表达量均显著低于IL-33刺激组,差异有统计学意义(P<0.05)。结论:Iloprost可能是ILC2s的负调节因子,在一定程度上可减轻小鼠肺部急性过敏性炎症反应。

Objective： To explore the effect of iloprost on ILC2 s in a mouse model of acute allergic airway inflammation induced by IL-33. Methods： C57 BL/6 mice aged 6-8 weeks were randomly divided into four groups： DMSO control group,IL-33 stimulation group,IL-33＋iloprost treament group and single iloprost group. Inflammatory cell infiltration in lung tissue was observed by HE staining,and mucus secretion was observed by PAS staining. The number of eosinophils（ EOS） and group 2 innate lymphoid cells（ ILC2 s） in bronchoalveolar lavage fluid（ BALF） and lung tissues was analyzed by flow cytometry. The cytokine levels of IL-5 and IL-13 were detected by ELISA. The expressions of IL-5,IL-13,GATA3 and ST2 at mRNA level were tested by real-time quantitative PCR. Results：（1）HE and PAS staining of lung sections revealed that a large number of inflammatory cells infiltrated and mucus secretion increased in the lung tissue sections of IL-33 simulation group. Besides,the number of eosinophils and ILC2 s,the levels of IL-5 and IL-13 and the expression of IL-5,IL-13,GATA3 and ST2 at mRNA levels were significantly higher than those in DMSO control group and single iloprost group,the difference was statistically significant（ P〈0. 05）.（2）It was confirmed that iloprost could significantly alleviate the inflammatory cells and mucus secretion in the lung tissue sections of IL-33 ＋ iloprost treatment group. In addition,iloprost could also inhibit the number of ILC2 s and eosinophils,reduce the levels of IL-5 and IL-13 and attenuate the expression of IL-5,IL-13,GATA3 and ST2 at mRNA level,compared with IL-33 model group（ P〈0. 05）. Conclusion： Iloprost may be a negative regulator of the ILC2 s,which can alleviate acute allergic airway inflammation in mice to some extent.